Comparison of a rebound and an applanation tonometer for measuring intraocular pressure in normal rabbits

Objective To compare intraocular pressure (IOP) measurements made on healthy adult rabbits without the effect of tranquilizers using the new applanation tonometer, Tono‐Pen Avia^®, and the rebound tonometer Tonovet^®. Methods Intraocular pressure was measured throughout the day (6:00, 9:00, 12:00, 15:00, and 18:00 h) in 38 adult New Zealand White rabbits (76 eyes). The animals were 20 males and 18 females, with a mean weight of 3.5 kg and an average age of 6 months. A complete ocular exam (including Schirmer tear test, fluorescein staining, slit‐lamp biomicroscopy, and direct ophthalmoscopy) was performed on all animals at the beginning of the trial. Rebound tonometry was performed, and after 10 min, anesthetic drops were instilled and applanation tonometry was carried out. IOP values obtained using the two techniques were analyzed statistically. Results The mean IOP was 9.51 ± 2.62 mmHg with Tonovet^®, and 15.44 ± 2.16 mmHg with the Tono‐Pen Avia^®. Significant differences between measurements with the two tonometers were observed (P < 0.001). The linear regression equation describing the relationship between the two tonometers was y = 0.4923x + 10.754 (y = Tonovet^® and x = Tono‐Pen Avia^®). High IOPs were recorded in the early measurements (6:00), but the average IOPs from both devices were statistically similar throughout the day (P = 0.086). The correlation coefficient was r² = 0.357. No significant difference in IOP regarding gender was observed. Conclusion The Tono‐Pen Avia^® recorded higher levels of IOP compared with the Tonovet^®. Early in the day, the IOP of rabbits was higher than later in the day, regardless of the tonometer used.     

Diversity of Paenibacillus durus strains isolated from soil and different plant rhizospheres evaluated by ARDRA and gyrB-RFLP analysis

Strains belonging to Paenibacillus durus isolated from the rhizosphere of various grasses and from bulk soil were previously divided into five phenotypic groups (A1–A5) based on the fermentation pattern of six carbohydrates (A1: sorbitol (+), A2: dulcitol and tagatose (+), A3: starch and glycogen (+), A4: starch, glycogen and d-arabitol (+) and A5: negative for these carbohydrates). This study aimed to assess whether plant types select for specific P. durus phenotypic groups. For that purpose, polymerase chain reaction-restriction fragment length polymorphism analysis of part of genes encoding 16S rRNA (ARDRA) and DNA gyrase subunit B (gyrB-RFLP) were used to produce genetic fingerprints. ARDRA and gyrB-RFLP data were clustered together to generate a dendrogram and two main clusters were observed. Cluster I showed a predominance of strains isolated from wheat, maize and sugarcane rhizospheres. Strains isolated from maize were distributed among the five patterns of carbohydrate metabolism, while strains isolated from sugarcane showed to be predominantly able to metabolize starch and glycogen. Neither sorbitol- nor arabitol-metabolizing strains were found in cluster II, which consisted of strains isolated from soil and from all plant species used. Our results suggest that the plants influenced the diversity of P. durus in their rhizospheres.     

Investigating phenotypic variability in Colletotrichum lindemuthianum populations

Colletotrichum lindemuthianum, causal agent of anthracnose in the common bean, has wide genetic variability. Differential bean cultivars and morphological and physiological characteristics were used to analyze 74 isolates of C. lindemuthianum collected in two counties in the state of Minas Gerais, Brazil. Six different races were found, with a predominance of race 65 at both locations. Isolates were classified according to their sensitivities to the fungicide thiophanate-methyl, normally used in the control of common bean anthracnose. In all, ≈10% of isolates were resistant to the fungicide in vitro. Characteristics such as indexes of mycelia growth rate, colony diameter, sporulation capacity, and percentage of germination demonstrated the high genetic variability of C. lindemuthianum. We also observed variation in conidial cytology. The conidia of most isolates showed septa formation after germination, in contrast to septa absence, previously reported in the literature. Sexual and asexual reproduction were evaluated for mechanisms that may contribute in the generation of variability in C. lindemuthianum. Conidial anastomosis tubes were commonly found, indicating that asexual reproduction can help increase variability in this species. Information from this study confirmed high variability in C. lindemuthianum and will guide future studies in basic knowledge and applied technologies.     

Biometrical, biological, biochemical and molecular characteristics of Meloidogyne incognita isolates and related species

Meloidogyne incognita is one of the most polyphagous species of root-knot nematodes occurring in Brazil and worldwide. Eight M. incognita isolates were studied, representing two enzymatic phenotypes (esterase and malate desydrogenase: I1/N1, I2/N1) and four cryptic Meloidogyne sp.1 (S2/N1) isolates, representing one cytological type (3n?=?40–46). Three M. hispanica isolates (Hi3/N1, 2n?=?32–36) and two of an atypical Meloidogyne sp.2 (S2a/N3, 3n?=?40–44) were included in this study for comparison. All isolates were tested with three M. incognita-specific molecular markers. The primer pairs B06F/R, miF/R and incK14F/R amplified three species-specific fragments of 1,200?bp, 955?bp and 399?bp, respectively for M. incognita and Meloidogyne sp.1 isolates. No amplification occurred in the M. hispanica and Meloidogyne sp.2 isolates, except with primers miF/R (1,650?bp). The genetic variability of the Meloidogyne spp. isolates was evaluated, using RAPD and ISSR markers. The phylogenetic analyses revealed two strongly supported monophyletic clades: clade I, consisting of M. hispanica and the atypical Meloidogyne sp.2 isolates, and clade II, clustering together all M. incognita and the Meloidogyne sp.1 isolates. Considering the biometrical, cytological and molecular approaches, it was possible to conclude that the isolates with three enzymatic phenotypes (I1/N1, I2/N1 and S2/N1) presented the characteristics described for M. incognita. Some correlations were detected between the isozymatic phenotypes and the tree topology (S2a/N3, Hi3/N1, I1/N1, S2/N1), but no strict correlation could be observed for the phenotype I2/N1 and one isolate of S2/N1. Morphologically, the Msp.2 isolates differ from M. incognita and M. hispanica by the female stylet features presenting straight cone tip and round pear shaped knobs, posteriorly sloping. The results of this study suggested that the Msp.2 isolates with phenotypes S2aN3 belong to a new or an unidentified species closely related to M. hispanica.     

Grafts of porcine small intestinal submucosa seeded with cultured homologous smooth muscle cells for bladder repair in dogs

Background

Due to numerous complications associated to gastrointestinal augmented cystoplasty, this study aimed to analyze the anatomic repair of the bladder of 10 female dogs using grafts of porcine small intestinal submucosa (SIS) seeded with cultured homologous smooth muscle cells, and compare them with the acellular SIS grafts.

Results

We assessed the possible side effects and complications of each type of graft by clinical examination, abdominal ultrasound and laboratory findings. Anatomic repair of neoformed bladder was assessed by histological staining for H/E and Masson''s Trichrome, analyzed with a Nikon Photomicroscope connected to the system of image analysis Image J.

Conclusions

We propose that SIS associated to homologous smooth cells can improve the quality of tissue repair, and consequently decrease the potential complications inherent to acellular SIS.     

Pseudomonas associated with Bursaphelenchus xylophilus,its insect vector and the host tree: A role in pine wilt disease?

In this study, we characterized the diversity of Pseudomonas associated with Bursaphelenchus xylophilus, its insect vector (Monochamus galloprovincialis) and its host (Pinus pinaster), by a culture‐independent approach using rpoD clone libraries. Clone libraries of Pseudomonas rpoD were obtained from B. xylophilus, M. galloprovincialis and infected P. pinaster. Most M. galloprovincialis and B. xylophilus sequences grouped together in the P. fluorescens group. Genes related to xenobiotics degradation and phenylacetate synthesis were present in the genomes of the type strains closely related to sequences retrieved from the nematode libraries. Results demonstrated that the nematode, during its life stages inside the tree, maintains a diverse Pseudomonas community that is closely related to the one associated with the insect vector. These bacteria might contribute to degradation of xenobiotics and tree weakening during the nematode tree infection.     

Invasive dynamics of the crayfish Procambarus clarkii (Girard, 1852) in the international section of the River Minho (NW of the Iberian Peninsula)

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